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一种基于合成螯合剂和绿色荧光蛋白的化学遗传指示剂,用于细胞内钠离子成像。

A chemigenetic indicator based on a synthetic chelator and a green fluorescent protein for imaging of intracellular sodium ions.

作者信息

Takeuchi Shiori, Imai Shosei, Terai Takuya, Campbell Robert E

机构信息

Department of Chemistry, Graduate School of Science, The University of Tokyo 7-3-1 Hongo Bunkyo-ku Tokyo 113-0033 Japan

CERVO Brain Research Center and Department of Biochemistry, Microbiology, and Bioinformatics, Université Laval, Québec Québec G1V 0A6 Canada.

出版信息

RSC Chem Biol. 2024 Dec 3;6(2):170-174. doi: 10.1039/d4cb00256c. eCollection 2025 Feb 5.

Abstract

A chemigenetic indicator with an affinity suitable for imaging of intracellular sodium ions (Na) in mammalian cells was developed. The indicator, based on a chimera of green fluorescent protein (GFP) and HaloTag labeled with a synthetic crown ether chelator, was produced by a combination of rational design and directed evolution. In mammalian cells the indicator exhibited an approximately 100% increase in excitation ratio when the cells were treated with 20 mM Na and an ionophore.

摘要

开发了一种化学遗传指示剂,其亲和力适合对哺乳动物细胞内的钠离子(Na)进行成像。该指示剂基于绿色荧光蛋白(GFP)和标记有合成冠醚螯合剂的HaloTag的嵌合体,通过合理设计和定向进化相结合的方式产生。在哺乳动物细胞中,当用20 mM Na和离子载体处理细胞时,该指示剂的激发比增加了约100%。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5288/11796374/4eafef3753da/d4cb00256c-f1.jpg

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